Figure 5 from Loss of Predicted Cell Adhesion Molecule MPZL3 Promotes EMT in Ovarian Cancer

MPZL3 knockdown leads to cell-cycle arrest and senescence-like phenot. A, Cell growth assay following knockdown of MPZL3 in OVCAR4 and OVCA433 cells. Cell growth (FluoReporter Blue Fluorometric dsDNA Quantitation) was normalized to day 1 (n = 3; two-way ANOVA group factor variance P < 0.0001; Dunnett multiple comparison test *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001). B, Semi-quantitative RT-PCR analysis of CCND1 transcript levels in OVCAR4 and OVCA433 cells following MPZL3 knockdown (n = 3; one-way ANOVA P < 0.0001; Dunnett multiple comparison test P values shown). C, CCND1 expression positively correlates with MPZL3 expression in TCGA ovarian serous adenocarcinoma specimens (mRNA z-scores; RNA-seq V2 RSEM). D, Cell-cycle analysis (propidium iodide flow cytometry) of OVCAR4 cells following MPZL3 knockdown at 9-hour timepoint from release (n = 3; two-way ANOVA group factor variance P < 0.0001; Dunnett multiple comparison test P values shown). E, Images of senescence-associated β-galactosidase (SA-β-gal) staining in OVCAR4 cells (scale bar, 200 μm). The corresponding quantification data are shown on the right (n = 3; one-way ANOVA P = 0.0029; Dunnett multiple comparison test P values shown). F, Heatmap of senescence-associated secretory phenotype markers and lamin B1 (LMNB1) following MPZL3 knockdown (OVCAR4 RNA-seq, z-scores). CCND1, cyclin D1; CI, confidence interval.